CN106974906A - Application of the andrographolide in the medicine for improving bleomycin antitumous effect is prepared - Google Patents
Application of the andrographolide in the medicine for improving bleomycin antitumous effect is prepared Download PDFInfo
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- A61K31/00—Medicinal preparations containing organic active ingredients
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Abstract
The present invention relates to application of the andrographolide in the medicine for improving bleomycin antitumous effect is prepared, this applies the weight percentage of andrographolide in described medicine to be 76.92% 86.96%.Medicine of the present invention is used cooperatively with bleomycin, life cycle, body weight, abdominal circumference diameter and the vigor of tumor model mouse can be significantly improved, the pulmonary fibrosis degree of tumor model mouse is substantially alleviated simultaneously, points out the pharmaceutical composition to mitigate the pulmonary fibrosis as caused by bleomycin while bleomycin antitumous effect is strengthened.
Description
Technical field
Match somebody with somebody product the present invention relates to medical, and in particular to the new medicinal usage of andrographolide.
Background technology
Andrographolide (Andrographolide, hereinafter referred to as Andro), natural plants Herba Andrographitis it is main effectively into
Part, with thermal detoxification of dispelling, the effect of anti-inflammatory analgetic, with viral infection of upper respiratory tract and dysentery there is special efficacy to bacillary,
It is described as natural antibiotics medicine.Experiment in vitro is proved：Breast cancer cellular synthesis of the Herba Andrographitis to culture has inhibitory action.
At present, proof andrographolide independent medication is not tested still has inhibitory action to tumour.Also still no andrographolide can increase
Strong bleomycin antitumor curative effect and the pertinent literature report for reducing its toxic side effect.
Chemotherapy is the mode for the treatment of tumour the most frequently used at present, and bleomycin (Bleomycin, hereinafter referred to as BLM) is one
Plant relatively common clinical chemotherapy medicine.BLM, a kind of glycopeptide antibiotic for coming from streptomyces verticillatus, broad-spectrum anti-cancer drug,
It is effective against various cancers such as lymthoma, orchioncus, the cancer of the esophagus and squamous cell carcinoma, lymthoma, orchioncus and pernicious
Pleural effusion etc..BLM is that it is antineoplastic while without the suppression of obvious marrow as the maximum advantage of clinical antitumor agents
Make and use, while the immunologic function to body does not have inhibitory action yet.However, the same with most of chemotherapeutics, BLM also has not
Hold the side effect ignored：BLM, which is used for a long time, can cause pneumonia sample symptom and pulmonary fibrosis symptom, thus bleomycin is used for a long time
It is also breakneck to treat tumour.Current most chemotherapeutics has different degrees of bone marrow suppression and immunosupress pair
Effect, thus as the chemotherapeutics BLM without this two shortcomings in clinical practice it is still very promising.But do not have also now
There are the technological means and active drug that can reduce bleomycin pulmonary fibrosis this toxic side effect.Therefore finding one kind can reduce
The medicine of BLM pulmonary fibrosis side effects is very urgent.
The content of the invention
The technical problem to be solved in the present invention is to provide the new application of andrographolide, i.e., andrographolide is in pharmacy
New application.
Specifically, new application of the above-mentioned andrographolide in pharmacy is that andrographolide improves rich next mould in preparation
Application in the medicine of plain antitumous effect.
In above-mentioned application, the weight percent content of andrographolide is 76.92%-86.96% in the medicine.
In above-mentioned application, described medicine is injection or common oral agents.
Pharmaceutical composition of the present invention can be prepared according to methods known in the art, i.e., by active ingredient and medically
Acceptable auxiliary material is mixed and made into conventional pharmaceutical preparation, such as tablet, capsule, parenteral solution, powder-injection or dripping pill.
The beneficial effects of the invention are as follows we are by using H22Cell (2 × 106Cells/ is only) ascites is made to kunming mice
Knurl model, gives various concentrations Andro solution, intraperitoneal injection BLM solution (15mg/kg) and administering drug combinations to control using gavage
Treat H22Ascites tumor mouse, so as to evaluate whether Andro can improve BLM tumor effects.Experiment one passes through Andro, BLM and two
Whether the apoptosis rates of Ascitic Tumor Cells proves Andro in influence of person's use in conjunction to mouse ascites volume and detection ascites
BLM antitumor action can be strengthened；Experiment two passes through the ascites tumor mouse that is crossed to Andro, BLM and both Combined Treatments
Lung tissue does histopathological examination to detect pulmonary fibrosis degree, to prove whether Andro has protective effect, i.e., can
Mitigate the pulmonary fibrosis of BLM inductions.Can Andro is observed by two experiments strengthen the effect of BLM treatment tumours and subtract
Light BLM causes the side effect of pulmonary fibrosis.
As a result show that Andro and BLM drug combinations have effectively taken on a new look the state of mind of ascites tumor mouse, reduce ascites
Abdominal circumference diameter, the ascites volume of knurl mouse, improve the apoptosis rate of mouse ascites oncocyte, significantly enhance the anti-of bleomycin
Tumor effect.Shown simultaneously by the observation to histotomy, Andro and BLM drug combinations can effectively improve BLM to lung's group
The pneumonia sample symptom and pulmonary fibrosis symptom weaved, have protective effect to lung tissue.It imply that Andro can improve BLM's
Antitumous effect, it is shown that it is preparing the good prospect in improving BLM antitumous effect medicines.
Description of test Andro and Andro enhancing BLM during BLM use in conjunction antitumor action in detail below,
Indicate beneficial effects of the present invention.Wherein Andro is the pulvis that this experiment teaching and research room is independently made.
One .Andro enhancings BLM antitumor action experiment.
1.1 experiment material
Male Kunming (KM) mouse (18-22g) is purchased from Guangdong Medical Lab Animal Center (certificate number SCXK2013-
0085) 98% andrographolide is purchased from the Sigma-Aldrich companies in the U.S., and it is limited that bleomycin is purchased from Japanese chemical drug
Company, H22 cells, obtained from American Type Culture Collection (Rockville, MD, USA), hyclone
(FBS) and the culture mediums of RPMI 1640 be purchased from Gibco (Grand Island, NY, US), phosphate buffered saline (PBS) (PBS), mould
Element and streptomysin are provided by Hyclone (Logan, Utah, US).
1.2 experiment packet
18~22g of SPF levels male KM mouse 10 is taken only to do normal group, intraperitoneal injection of saline, remaining 84 heros
Property KM mouse use H22Cell (2 × 106Cells/ is only) make ascites tumor model to mouse peritoneal injection method.80 are selected after 5 days
The successful ascites tumor mouse of modeling is randomly divided into agent in model control group, BLM groups, Andro low dose groups (Andro-L), Andro
Amount group (Andro-M), Andro high doses group (Andro-H), Andro-L+BLM groups, Andro-M+BLM groups, Andro-H+BLM
Group.
1.3 experimental method
Start to be administered after packet, BLM groups intraperitoneal injection BLM solution (15mg/kg, with normal saline), Andro-L,
Andro-M, Andro-H group respectively gavage Andro solution (prepared with CMC-Na, concentration be followed successively by 25mg/kg, 50mg/kg,
100mg/kg), BLM solution (15mg/kg) connection is injected intraperitoneally in Andro-L+BLM, Andro-M+BLM, Andro-H+BLM group respectively
Gavage Andro solution (concentration is followed successively by 25mg/kg, 50mg/kg, 100mg/kg) is closed, normal group and model control group gavage are given
Medicine CMC-Na, once a day, successive administration 7 days, last time administration are put to death after 24 hours, collect ascites and lung tissue.
1.4 Testing index and detection method
1.4.1 the general state of each group mouse is observed
In experimentation, change of observation each group mouse diet, hair, the state of mind and build etc. daily.
1.4.2 observation each group mouse weight and abdominal circumference change and death condition.
From inoculation, daily 9 groups of mouse weights of periodic monitor, abdominal circumference and death state are simultaneously recorded, and are as a result united
Processing is counted, form is drawn.
1.4.3 the apoptosis rate of the Ascitic Tumor Cells in flow cytomery mouse ascites
Mouse is put to death, ascites is gathered, at 4 DEG C, centrifuges 3 minutes, is then washed with PBS 2 times under conditions of 3000rpm,
Dye and detected by Countstar automatic cell counters, harvesting using 0.2% trypan blue analysis cell viability.It is all
The cell of group is centrifuged and washed twice with ice-cold PBS, is then resuspended in combination buffer.Finally, cell is subjected to film connection
Albumen V-FITC- propidium iodides (PI) double dyeing simultaneously pass through flow cytomery apoptosis rate.Determination data is thin by streaming
The software that born of the same parents' instrument is configured carries out data processing, and the early apoptosis rate represented with bottom right adds the middle and later periods apoptosis rate that upper right is represented
Summation represent natural death of cerebral cells rate.
1.5 statistical analysis
It is poor between statistical procedures, each group that all data are carried out using for Windows statistical analysis softwares of SPSS 17.0
It is different compare use variance analysis, with P<0.05 is that difference has conspicuousness.
1.6 experimental result
1.6.1 each group mouse animation
The low middle high group mouse weight of blank control group and andrographolide independent medication increases sharply after being inoculated with 5 days,
One's spirits are drooping depressed, dull few dynamic, instability of gait is then engendered, reactivity is poor, and water inlet feed is reduced, and late period belly is swollen
It is grand, in dyscrasia state.Andrographolide combines group with bleomycin except low dose group increased weight is very fast, it is One's spirits are drooping it
Remaining outer two groups of increased weight are slower, and spirit is preferably.
1.6.2 the body weight of each group mouse, abdominal circumference and ascites volume situation
Compared with model group, BLM groups, Andro-L+BLM groups, the body weight of Andro-M+BLM groups and Andro-H+BLM groups and
Abdominal circumference has significant difference (P<0.01), the body weight and abdominal circumference of Andro-L groups, Andro-M groups and Andro-H groups do not show
Write sex differernce (P>0.05).Compared with BLM groups, Andro-L+BLM groups (P<0.05), Andro-M+BLM groups and Andro-H+BLM
Group (P<0.01) abdominal circumference has significant difference.Refer to table 1 below, table 2：
Table 1：Experiment mice body weight (g) changes table
a：P ＜ 0.05, aa：P ＜ 0.01, are compared with blank group；b：P ＜ 0.05, bb：P ＜ 0.01, are compared with model group；
c：P ＜ 0.05, are compared with BLM groups
Table 2：Experiment mice abdominal circumference diameter (mm) table
a：P ＜ 0.05, aa：P ＜ 0.01, are compared with blank control group；b：P ＜ 0.05, bb：P ＜ 0.01, with model group ratio
Compared with；c：P ＜ 0.05, cc：P ＜ 0.01, are compared with BLM groups
Compared with model group, BLM groups (P ＜ 0.05), Andro-L+BLM groups, Andro-M+BLM groups and Andro-H+BLM
The abdominal circumference of group (P ＜ 0.01) has significant difference.The abdominal circumference of Andro-M+BLM groups and Andro-H+BLM groups compared with BLM groups
There is significant difference (P ＜ 0.05).Refer to Fig. 1.
As a result show, BLM and Andro drug combinations can significantly reduce body weight, abdominal circumference and the ascites volume of ascites tumor mouse.
1.6.3 the apoptosis rate of the Ascitic Tumor Cells of each group mouse
Compared with model group, BLM groups and BLM+Andro group Ascitic Tumor Cells apoptosis rates substantially rise (P<0.01) it is and small
The apoptosis rate of mouse ascites fluid oncocyte can be improved with the increase of Andro dosage；Compared with BLM groups, the abdomen of Andro-H+BLM groups
Hydatoncus apoptosis rate is significantly improved (P<0.05).As a result show, BLM and Andro drug combinations can significantly improve small
The apoptosis rate of mouse ascites fluid oncocyte.Refer to Fig. 2.
Two, andrographolides mitigate pneumonia sample symptom and the pulmonary fibrosis experiment of BLM inductions
2.1 experiment packet
Strengthen 1.2 in BLM antitumor action experiment with Andro.
2.2 experimental method
The lung group of 1.3 Collection and conservations in the antitumor action experiment that lung tissue used strengthens BLM from Andro
Knit.The lung tissue sample of mouse is quickly cleaned with physiological saline, lung tissue is fixed with 10% formalin, lung tissue is with often
Rule organizational technology processes and embedded with paraffin.5mm slabs will be cut into the sample of FFPE, be placed on slide
On, dyed with h and E, carry out pathological observation；Collagen deposition is contaminated with Masson trichrome stainings
Color, is identified by characteristic blue.Injury of lungs is evaluated according to following standard：Alveolar redness degree, the thickness of alveolar wall and
The hyperplasia degree of connective tissue, collagen deposition and inflammation in the increase degree of alveolar, essence.Histopathological lesions are with 0-4's
Scale (0 represents normal, does not damage, 1 is 25%, minor injury, and 2 be 25-50%, moderate lesion, and 3 be 50-75%,
Severe injury, 4 be 75-100%, is extremely damaged).Overall merit is carried out to injury of lungs according to the synthesis of the score of 4 standards.Root
Judge whether Andro can mitigate pneumonia sample symptom and the pulmonary fibrosis of BLM inductions according to the degree of injury of lungs.
2.3 statistical analysis
It is poor between statistical procedures, each group that all data are carried out using for Windows statistical analysis softwares of SPSS 17.0
It is different compare use variance analysis, with P<0.05 is that difference has conspicuousness.
2.4 experimental result
As a result show that lung tissue's section of normal group (Fig. 3 A1, B1) and model group (Fig. 3 A2, B2) becomes without obvious pathology
Change, it may be observed that normal bronchus and alveolar and a small amount of collagen deposition；Lung tissue's section of BLM groups (Fig. 3 A3, B3)
Substantially, alveolar wall is thickening, and inflammatory infiltration occur in alveolar and the congestion of blood vessel, perivascular cell and bronchial epithelial cells for pathological change,
Pulmonary fibrosis and collagen deposition, serious lung injury degree are high；Andro-L+BLM groups (Fig. 3 A4, B4), Andro-M+BLM groups (figure
3A5, B5) and the pathological condition of Andro-H+BLM groups (Fig. 3 A6, B6) progressively weaken compared with BLM groups, serious lung injury degree is successively
Decline.Fig. 3 block diagram is shown:Compared with normal group, the serious lung injury degree highest (P of BLM groups<0.01)；With BLM group phases
Than the serious lung injury degree of Andro-L+BLM groups, Andro-M+BLM groups and Andro-H+BLM groups has substantially reduction (P<
, and serious lung injury degree weakens with the increase of Andro dosage 0.01).BLM inductions can be mitigated by demonstrating Andro
Pneumonia sample symptom and pulmonary fibrosis.
Brief description of the drawings
Fig. 1 is BLM and the mouse ascites volume histograms of BLM and Andro drug combination groups.
Fig. 2 is the design sketch of the influence of BLM and Andro and BLM drug combinations to mouse ascites apoptosis of tumor rate, its
In, A figures are flow cytometer result figure, and B figures are apoptosis of tumor cells rate block diagram.
Fig. 3 is the design sketch of the influence of BLM and Andro and BLM drug combinations to mouse lung tissue, wherein, top is
Lung tissue SABC microphoto, bottom is injury of lungs grade block diagram.
Embodiment
According to known method in medical industry, using Andro as BLM synergy attenuation medicine, with one
Kind or a variety of pharmaceutically acceptable carriers or excipient are mixed, and the pharmaceutical composition of different dosage forms is made.Including oral medicine
Agent or injection medicament, the medicinal preparation for oral administration can be dripping pill, soft capsule etc.；The injection medicament can be emulsion for injection, note
It can be emulsion used for intravenous injection, intramuscular injection with emulsion or hypodermic injection emulsion to penetrate emulsion.
Following examples are not offered as that present invention is limited only by the embodiment.
Embodiment 1 (self-emulsifying soft capsule)
Andrographolide 5g, adds 500ml oil phase (ethyl oleate 5ml), emulsifying agent (Tween-80 27ml) and helps breast
Agent (n-butanol 18ml), it is dissolved with ultrasonic wave, and the solution of gained is pressed into flexible glue with gelatin through automatic rotation rolling capsule machine
Capsule, 5mg/, you can be orally united and applied in oncotherapy with BLM.
Embodiment 2 (dripping pill)
Andrographolide 150g, takes the ferment 6000 of poly- second two, the Hard Macrogol of equivalent, is well mixed, and heating melts, plus
Enter andrographolide, mix, dripping pelletization, film coating, you can be orally united and applied in oncotherapy with BLM.
Claims (3)
1. application of the andrographolide in the medicine for improving bleomycin antitumous effect is prepared.
2. application according to claim 1, it is characterised in that the weight percent content of andrographolide in the medicine
For 76.92%-86.96%.
3. application according to claim 2, it is characterised in that described medicine is injection or common oral agents.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018177301A1 (en) * | 2017-04-01 | 2018-10-04 | 郑州大学 | 15-idene-14-deoxy-11,12-dehydroandrographolide derivative and application thereof in preparing anti-fibrosis drugs |
| CN109730990A (en) * | 2018-03-02 | 2019-05-10 | 郑州大学 | Application of the 15- benzyl subunit -14- deoxidation -11,12- andrographolide in anti-fibrosis medicine |
| CN112480097A (en) * | 2020-11-26 | 2021-03-12 | 汕头大学医学院 | Inhibitor of targeting ETS structural domain protein and application thereof |
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| CN102702147A (en) * | 2012-06-18 | 2012-10-03 | 辽宁利锋科技开发有限公司 | Andrographolide analogue and application of andrographolide analogue to treatment |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102702147A (en) * | 2012-06-18 | 2012-10-03 | 辽宁利锋科技开发有限公司 | Andrographolide analogue and application of andrographolide analogue to treatment |
Non-Patent Citations (3)
| Title |
|---|
| HUIZHEN GUO等: "Enhanced anti-tumor activity and reduced toxicity by combination andrographolide and bleomycin in ascitic tumor-bearing mice", 《EUROPEAN JOURNAL OF PHARMACOLOGY》 * |
| TAO ZHU等: "Protective role of andrographolide in bleomycin-induced pulmonary fibrosis in mice", 《INT. J. MOL. SCI.》 * |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018177301A1 (en) * | 2017-04-01 | 2018-10-04 | 郑州大学 | 15-idene-14-deoxy-11,12-dehydroandrographolide derivative and application thereof in preparing anti-fibrosis drugs |
| CN109730990A (en) * | 2018-03-02 | 2019-05-10 | 郑州大学 | Application of the 15- benzyl subunit -14- deoxidation -11,12- andrographolide in anti-fibrosis medicine |
| CN109730990B (en) * | 2018-03-02 | 2021-07-02 | 郑州大学 | Application of 15-benzylidene-14-deoxy-11, 12-dehydroandrographolide derivative in anti-fibrosis drugs |
| CN112480097A (en) * | 2020-11-26 | 2021-03-12 | 汕头大学医学院 | Inhibitor of targeting ETS structural domain protein and application thereof |
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Application publication date: 20170725 |